Journal of Biomolecular NMR

, Volume 53, Issue 2, pp 139–148

4D Non-uniformly sampled HCBCACON and 1J(NCα)-selective HCBCANCO experiments for the sequential assignment and chemical shift analysis of intrinsically disordered proteins

  • Jiří Nováček
  • Noam Y. Haba
  • Jordan H. Chill
  • Lukáš Žídek
  • Vladimír Sklenář
Article

DOI: 10.1007/s10858-012-9631-8

Cite this article as:
Nováček, J., Haba, N.Y., Chill, J.H. et al. J Biomol NMR (2012) 53: 139. doi:10.1007/s10858-012-9631-8

Abstract

A pair of 4D NMR experiments for the backbone assignment of disordered proteins is presented. The experiments exploit 13C direct detection and non-uniform sampling of the indirectly detected dimensions, and provide correlations of the aliphatic proton (Hα, and Hβ) and carbon (Cα, Cβ) resonance frequencies to the protein backbone. Thus, all the chemical shifts regularly used to map the transient secondary structure motifs in the intrinsically disordered proteins (Hα, Cα, Cβ, C′, and N) can be extracted from each spectrum. Compared to the commonly used assignment strategy based on matching the Cα and Cβ chemical shifts, inclusion of the Hα and Hβ provides up to three extra resonance frequencies that decrease the chance of ambiguous assignment. The experiments were successfully applied to the original assignment of a 12.8 kDa intrinsically disordered protein having a high content of proline residues (26 %) in the sequence.

Keywords

Intrinsically disordered proteins Non-uniform sampling 13C detection Chemical shifts Residual secondary structure Prolines assignment 

Supplementary material

10858_2012_9631_MOESM1_ESM.pdf (179 kb)
PDF (180 KB)

Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Jiří Nováček
    • 1
  • Noam Y. Haba
    • 2
  • Jordan H. Chill
    • 2
  • Lukáš Žídek
    • 1
  • Vladimír Sklenář
    • 1
  1. 1.Faculty of Science, NCBR, and CEITECMasaryk UniversityBrnoCzech Republic
  2. 2.Department of ChemistryBar Ilan UniversityRamat GanIsrael

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